CRISPR-Cas9-based genome editing enables the rapid genetic manipulation of any genomic locus without the need for gene targeting by homologous recombination. Here we describe a conditional transgenic approach that allows temporal control of CRISPR-Cas9 activity for inducible genome editing in adult mice. We show that doxycycline-regulated Cas9 induction enables widespread gene disruption in multiple tissues and that limiting the duration of Cas9 expression or using a Cas9D10A (Cas9n) variant can regulate the frequency and size of target gene modifications, respectively. Further, we show that this inducible CRISPR (iCRISPR) system can be used effectively to create biallelic mutation in multiple target loci and, thus, provides a flexible and fast platform to study loss-of-function phenotypes in vivo.
Inducible in vivo genome editing with CRISPR/Cas9
L. Dow,Jonathan Fisher,K. O’Rourke,A. Muley,Edward R. Kastenhuber,Geulah Livshits,D. Tschaharganeh,N. Socci,S. Lowe
Published 2015 in Nature Biotechnology
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- Publication year
2015
- Venue
Nature Biotechnology
- Publication date
2015-02-10
- Fields of study
Biology, Medicine
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Semantic Scholar, PubMed
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